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    • 2. 发明申请
    • LENTIVIRAL VECTOR EXPRESSING MEMBRANE-ANCHORED OR SECRETED ANTIBODY
    • 表达载体表达的膜载体或分泌的抗体
    • WO2017005923A1
    • 2017-01-12
    • PCT/EP2016/066349
    • 2016-07-08
    • INSTITUT NATIONAL DE LA SANTE ET DE LA RECHERCHE MEDICALE (INSERM)UNIVERSITÉ CLAUDE BERNARD LYON 1CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUEECOLE NORMALE SUPERIEURE DE LYON
    • FUSIL, FlorianeVERHOEYEN, ElsDEFRANCE, ThierryCOSSET, François-Loïc
    • C12N15/86C07K16/10A61K39/29
    • C12N15/86A61K39/29A61K2039/505A61K2039/5256C07K16/00C07K16/109C07K2317/10C07K2317/76C07K2319/03C12N2740/15043C12N2810/6054C12N2830/008C12N2830/20C12N2830/42C12N2830/48C12N2830/50
    • The invention concerns a multicistronic nucleic acid, in particular an isolated multicistronic nucleic acid, comprising: A) a sequence comprising successively: A1) a sequence encoding the light chain variable domain of an antibody of interest, fused in the frame with A2) a sequence encoding the constant region of the light chain of an immunoglobulin Ig; and B) a sequence comprising successively: B1) a sequence encoding the heavy chain variable domain of said antibody of interest, fused in the frame with B2) a sequence encoding the constant regions of the heavy chain of an immunoglobulin Ig' in secretory form; B3) an intronic sequence of the gene of the heavy chain of said immunoglobulin Ig', said intronic sequence comprising an internal 5' splice site enabling the splicing of said intronic sequence B3) and a secretory-specific poly(A) (p AS) signal from the 3' terminal exon of said gene; B4) a sequence, in frame with sequence B1), encoding the transmembrane and cytoplasmic domains M1 and M2 of the immunoglobulin Ig' BCR, wherein said sequence B4) comprises, between the coding sequences of the M1 and M2 domains, an intronic sequence containing a splice site enabling the splicing of said intronic sequence between the M1 and M2 domains coding sequences; and B5) a membrane-anchored specific poly(A) signal (p AM), after the stop codon of the M2 domain, wherein the multicistronic nucleic acid enables the co-expression of the sequences A and B into separate proteins.
    • 本发明涉及多顺反子核酸,特别是分离的多顺反子核酸,其包含:A)连续包含的序列:A1)编码感兴趣抗体的轻链可变结构域的序列,其融合在框架中A2)序列 编码免疫球蛋白Ig的轻链的恒定区; 和B)连续包含的序列:B1)编码所述感兴趣抗体的重链可变结构域的序列,其与所述框架融合在一起; B2)编码分泌形式的免疫球蛋白Ig'的重链恒定区的序列; B3)所述免疫球蛋白Ig'的重链基因的内含子序列,所述内含子序列包含能够剪接所述内含子序列B3)和分泌特异性聚(A)(p AS)的内部5'剪接位点, 来自所述基因的3'末端外显子的信号; B4)编码免疫球蛋白Ig'BCR的跨膜和胞质结构域M1和M2的序列,其中序列B1),其中所述序列B4)在M1和M2结构域的编码序列之间包含含有 所述剪接位点能够在M1和M2结构域编码序列之间剪接所述内含子序列; 和B5)M2结构域的终止密码子之后的膜锚定的特异性聚(A)信号(pAM),其中所述多顺反子核酸能够使序列A和B共同表达成分离的蛋白质。